Figure 2.

miR-122-dependent and miR-122-independent replication systems. (A) Cartoon diagram of Full-length (FL) Rluc HCV RNA (top) and depiction of endogenous miR-122 binding to the 5′ terminus of the 5′ UTR of FL viral RNA (bottom). (B) Diagram of subgenomic replicon (SGR) FLuc S1+S2:p3 HCV RNA (top) and depiction of miR-122-independent (miRControl, bottom left) and miR-122-dependent (miR-122p3, bottom right) replication systems. Since Huh-7 and Huh-7.5 cells endogenously express miR-122, endogenous miR-122 binding is abolished by introduction of point mutations in both of the miR-122 binding sites (S1+S2p3, indicated in red). miRControl (where the entire seed region of the miRNA is mutated, indicated in red) or miR-122p3 molecules (containing a compensatory mutation at position 3, indicated in red) are used for miR-122-independent and miR-122-dependent replication, respectively.